Celik Yamaci Merve, Pamukcu Ceren, Erdemgil Yigit, Atik Ahmet Emin, Keles Zeynep Zulfiye Yildirim, Can Ozge
Department of Medical Biotechnology, Institute of Health Sciences, Acibadem Mehmet Ali Aydinlar University, 34638 Istanbul, Türkiye.
Biotechnology Group, Turgut Ilaclari A.S., 41400 Kocaeli, Türkiye.
Pharmaceuticals (Basel). 2025 Apr 16;18(4):579. doi: 10.3390/ph18040579.
: The stress testing of biotherapeutic products is a critical component of drug development, enabling the assessment of stability, biosimilarity, and degradation pathways. Subjecting biosimilar monoclonal antibodies to controlled stress conditions yields essential insights into their structural and functional integrity, informing formulation optimization and mitigating risks before clinical trials. In this study, biosimilar products were comprehensively characterized and compared with originator products under forced degradation. The aim was to expose the products to different stress conditions such as oxidative, pH, thermal, freeze/thaw, and agitation. The products were then tested at defined time points using validated analytical methods. : This study employed size-exclusion chromatography to detect aggregated forms. Isoelectric focusing characterized protein charge variants (e.g., acidic/basic isoforms) from post-translational modifications, while capillary electrophoresis quantified product-related impurities (aggregates and fragments). In addition, a complement assay was used to determine the efficacy and potency under specific stress conditions. : Our findings showed that biosimilar and originator products exhibited similar degradation profiles. The biosimilar monoclonal antibody was found to be analytically similar to the originator product in terms of critical parameters related to efficacy and safety under various stress conditions such as aggregation profile, biological activity, and charge variant distribution. : Forced degradation studies facilitated the comprehensive and well-validated characterization of the structure and biological activity of biosimilar monoclonal antibody products.
生物治疗产品的应力测试是药物开发的关键组成部分，有助于评估稳定性、生物相似性和降解途径。使生物相似性单克隆抗体处于可控的应力条件下，能深入了解其结构和功能完整性，为制剂优化提供依据，并在临床试验前降低风险。在本研究中，对生物相似性产品进行了全面表征，并与强制降解条件下的原研产品进行了比较。目的是使产品暴露于不同的应力条件下，如氧化、pH值、热、冻融和搅拌。然后在规定的时间点使用经过验证的分析方法对产品进行测试。本研究采用尺寸排阻色谱法检测聚集形式。等电聚焦用于表征翻译后修饰产生的蛋白质电荷变体（如酸性/碱性异构体），而毛细管电泳则对与产品相关的杂质（聚集体和片段）进行定量。此外，还使用了补体测定法来确定特定应力条件下的效力和效能。我们的研究结果表明，生物相似性产品和原研产品具有相似的降解曲线。发现在各种应力条件下，如聚集情况、生物活性和电荷变体分布等与效力和安全性相关的关键参数方面，生物相似性单克隆抗体在分析上与原研产品相似。强制降解研究有助于对生物相似性单克隆抗体产品的结构和生物活性进行全面且经过充分验证的表征。
